Optimized protocols for efficient DNA extraction from leaves, stems, flowers, and seeds of Cannabis sativa L.

Abstract

Cannabis (Cannabis sativa L.) is becoming an increasingly important crop due to its applications in medicine, industry, and agriculture. Its complex genetics, coupled with varying legal statuses and the demand for high-quality varieties, have made reliable methods essential for the identification, classification, and improvement of cannabis plants. DNA extraction is a key technique enabling such research, which serves as the foundation for various molecular biology applications. The objetive of this study was to evalaute the effciacy of four different DNA extraction methods, using fresh samples of leaves, stems, flowers and seeds of cannabis.DNA extraction was performed in triplicate following four methods: saline precipitation with sodium dodecyl sulfate (SDS), potassium acetate precipitation (KA), DNAzol^TM reagent, and the commercial kit DNeasy Mini^TM from Qiagen (KC, control). The concentration, purity, and integrity of the extracted DNA were evaluated using spectrophotometry and agarose gel electrophoresis. Furthermore, the efficiency of DNA amplification was assessed through polymerase chain reaction (PCR). The results indicated that the KA extraction method yielded samples with higher DNA concentrations, whereas the SDS method consistently yielded DNA of higher purity across all four tissue types. The use of the KC and SDS methodologies for DNA extraction facilitated the full PCR amplification of all four tissues. In contrast, the DNAzol method achieved 100% amplification solely in the leaf and stem samples. Consequently, the SDS and DNAzol methodologies offer viable alternatives to the KC method, proving to be equally effective while being less labour-intensive and more economical for large-scale DNA extractions.